I got mcf-7 since 48 in 175 ml flask is confluent , so i spilit them in 5 flasks (25*50) ml
The appearance of cell using microscope is very well and cell count very rich.
After 48 hrs
The cells were stressed with square shape and i do not know what can i do, and what is wrong
Hello, Alaa Abdullah!
Cells feel uncomfortable when they are under stress, when they don't like something.
This can be a change in cultivation conditions: another medium, low serum concentration, high concentration of antibiotics, altered pH of the medium, contamination with microorganisms, fungi, lack or excess of CO2.
Also, some cells are very sensitive to the substrate, i. e. grow only on flasks or plates with high adhesion. In your case, you check the brand name (manufacturer) of your culture flasks. Maybe the 175 cm flasks were from a different manufacturer and with different plastic adhesive properties than the 25 cm flasks.
The typical culture medium for MCF-7 is alpha MEM (EMEM alpha modification)+ 10% FBS +insulin.
Alexandr Chernov
What about DMEM low glucose does it cause stress state?
and the suitable incubation time to trypsinized the cells
could you provide me with your cell culture protochool and the tricky steps
Dear Abudullah,
Many researchers have already changed the culture method. Probably your MCF-7 cells have been cultured in DMEM when you started the culture. However, there should be a certain reason to pick the suitable culture medium at the first time when the cells were established. So if you lose the way, you should go back to the start. Also I wrote about insulin. Do you add the insulin for the culture medium routinely? I thought the insulin might be the reason why MCF-7 was stressed.
If you add the insulin routinely, sorry I have no idea, just try the original culture method.
Good Luck,
- Badges
- Science topic
- Similar topics
- Agriculture
- Horticulture
- Seeds